Although clinical evidence suggests that nonalcoholic fatty liver disease is an established major risk factor for heart failure, it remains unexplored whether sleep disorder-caused hepatic damage contributes to the development of cardiovascular disease (CVD). Here, our findings revealed that sleep fragmentation (SF) displayed notable hepatic detrimental phenotypes, including steatosis and oxidative damage, along with significant abnormalities in cardiac structure and function. All these pathological changes persisted even after sleep recovery for 2 consecutive weeks or more, displaying memory properties. Mechanistically, persistent higher expression of nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) in the liver was the key initiator of SF-accelerated damage phenotypes. SF epigenetically controlled the acetylation of histone H3 lysine 27 (H3K27ac) enrichment at the Nox4 promoter and markedly increased Nox4 expression in liver even after sleep recovery. Moreover, fine coordination of the circadian clock and hepatic damage was strictly controlled by BMAL1-dependent Sirtuin 1 (Sirt1) transcription after circadian misalignment. Accordingly, genetic manipulation of liver-specific Nox4 or Sirt1, along with pharmacological intervention targeting NOX4 (GLX351322) or SIRT1 (Resveratrol), could effectively erase the epigenetic modification of Nox4 by reducing the H3K27ac level and ameliorate the progression of liver pathology, thereby counteracting SF-evoked sustained CVD. Collectively, our findings may pave the way for strategies to mitigate myocardial injury from persistent hepatic detrimental memory in diabetic patients.

A major obstacle in type 2 diabetes mellitus (T2DM) is sleep fragmentation (SF), which negatively affects testicular function. However, the underlying mechanisms remain to be elucidated. In this study, we demonstrate that SF induces testicular damage through a mechanism involving lipid metabolism, specifically mediated by melatonin (MEL) receptor 1a (MT1). T2DM mice with SF intervention displayed several deleterious phenotypes such as apoptosis, deregulated lipid metabolism, and impaired testicular function. Unexpectedly, sleep recovery (SR) for 2 consecutive weeks could not completely abrogate SF's detrimental effects on lipid deposition and testicular function. Interestingly, MEL and MT1 agonist 2-iodomelatonin (2IM) effectively improved lipid homeostasis, highlighting MEL/2IM as a promising therapeutic drug for SF-trigged testicular damage. Mechanistically, MEL and 2IM activated FGFR1 and sequentially restrained the crosstalk and physical interaction between TAB1 and TAK1, which ultimately suppressed the phosphorylation of TAK1 to block lipid deposition and cell apoptosis caused by SF. The ameliorating effect of MEL/2IM was overtly nullified in Fgfr1 knockout (Fgfr1-KO ^+/- ) diabetic mice. Meanwhile, testicular-specific overexpression of Tak1 abolished the protective effect of FGF1^mut on diabetic mouse testis. Our findings offer valuable insights into the molecular mechanisms underlying the testicular pathogenesis associated with SF and propose a novel therapeutic approach for addressing male infertility in T2DM.

OBJECTIVES: To explore the active components that mediate the therapeutic effect of Centella asiatica on psoriasis and their therapeutic mechanisms. METHODS: TCMSP, TCMIP, PharmMapper, Swiss Target Prediction, GeneCards, OMIM and TTD databases were searched for the compounds in Centella asiatica and their targets and the disease targets of psoriasis. A drug-active component-target network and the protein-protein interaction network were constructed, and DAVID database was used for pathway enrichment analysis. In a RAW264.7 macrophage model of LPS-induced inflammation, the anti-inflammatory effect of 7.5, 15, 30, and 60 μmol/L quercetin, asiaticoside, and asiatic acid, which were identified as the main active components in Centella asiatica, were tested by measuring cellular production of NO, TNF‑α and IL-6 using Griess method and ELISA and by detecting mRNA expressions of IL-23, IL-17A, TNF-α and IL-6 and protein expressions of p-STAT3 (Tyr705) and p-STAT3 (Ser727) with RT-qPCR and Western blotting. RESULTS: A total of 139 targets of Centella asiatica and 4604 targets of psoriasis were obtained, and among them CASP3, EGFR, PTGS2, and ESR1 were identified as the core targets. KEGG analysis suggested that quercetin, asiaticoside, and asiatic acid in Centella asiatica were involved in cancer and IL-17 and MAPK signaling pathways. In the RAW264.7 macrophage model of inflammation, treatment with quercetin significantly reduced cellular production of NO, TNF‑α and IL-6, and lowered mRNA expressions of IL-23, IL-17A, TNF‑α and IL-6 and protein expressions of p-STAT3 (Tyr705) and p-STAT3 (Ser727). CONCLUSIONS Quercetin, asiaticoside and asiatic acid are the main active components in Centella asiatica to mediate the therapeutic effect against psoriasis, and quercetin in particular is capable of suppressing cellular production of NO, TNF‑α and IL-6 and regulating the IL-23/IL-17A inflammatory axis by mediating STAT3 phosphorylation to inhibit inflammatory response.
Quercetin/pharmacology* ; Psoriasis/metabolism* ; STAT3 Transcription Factor/metabolism* ; Mice ; Animals ; Centella/chemistry* ; Triterpenes/pharmacology* ; Phosphorylation ; Interleukin-17/metabolism* ; Interleukin-23/metabolism* ; RAW 264.7 Cells ; Pentacyclic Triterpenes/pharmacology* ; Macrophages/drug effects* ; Signal Transduction ; Plant Extracts

Senecavirus A (SVA) is a major viral pathogen causing disease in pigs, and effective monitoring of SVA infection is critical for disease control. In this study, we aimed to develop a reliable ELISA method for rapidly detecting neutralizing antibodies against SVA. We used HEK293F cells to express an SVA-specific porcine Fab antibody and verified the biological activity of the Fab antibody by indirect ELISA, immunofluorescence assay, virus neutralization test, and Western blotting. The Fab antibody was biotinylated and used as a competitive antibody to establish a competitive ELISA (C-ELISA) for detecting neutralizing antibodies against SVA. We then evaluated the C-ELISA in terms of sensitivity, specificity, repeatability, and result agreement rate with the VNT. The results showed that we successfully prepared an SVA-specific porcine Fab antibody, which showed high affinity for SVA. We named this antibody 1M33Fab and designated it as Bio-1M33Fab after biotin labeling. The assay conditions were optimized as follows: the coating concentration of SVA particles being 1 μg/mL, the working concentration of Bio-1M33Fab being 0.5 μg/mL, the optimal serum dilution of 1:10, and the optimal dilution of enzyme-labeled avidin being 1:30 000. At a percent inhibition (PI) of 47%, the assay demonstrated the highest sensitivity (96.88%) and specificity (100%), with no cross-reactivity observed with the positive sera of major porcine viral diseases. The intra-assay coefficient of variation ranged from 1.12% to 7.34%, while the inter-assay coefficient of variation ranged from 1.10% to 8.97%, indicating good repeatability. In the detection of 224 clinical pig serum samples, C-ELISA and VNT showed a result agreement rate of 93.75%. In conclusion, we successfully develop a C-ELISA method for detecting neutralizing antibodies against SVA by using a porcine-derived Fab antibody, which lays a foundation for the development of detection kits.
Animals ; Swine ; Antibodies, Neutralizing/immunology* ; Enzyme-Linked Immunosorbent Assay/methods* ; Immunoglobulin Fab Fragments/immunology* ; Antibodies, Viral/immunology* ; Picornaviridae/immunology* ; Humans ; HEK293 Cells ; Swine Diseases/diagnosis* ; Picornaviridae Infections/diagnosis*

OBJECTIVE To establish the fingerprint of Lianhua Qingwen capsule(LHQW) with the HPLC method, and to evaluate the quality by combining with chemical pattern recognition method. METHODS The chromatographic separation was performed on an Agilent Zorbax SB-C18(250 mm×4.6 mm, 5 μm) column with 0.1% phosphoric acid(A)-acetonitrile(B) as the mobile phase for the gradient elution. The detection wavelength was set at 207 nm. The flow rate was set at 1.0 mL·min–1 and the column temperature was 30 ℃. Similarity evaluation, hierarchical clustering analysis(HCA), radar plot analysis, principal component analysis(PCA), and orthogonal partial least-squares discrimination analysis(OPLS-DA) were used for the further assessment of 13 batches of LHQW samples. RESULTS The fingerprint of LHQW was established with 40 common peaks, in which 10 common peaks were identified, and the similarities of 13 batches of LHQW samples were 0.947–1.000. By applying chemical pattern recognition methods such as HCA, and OPLS-DA, 13 batches of samples were classified into three clusters, and the results of classification were correlated with the production date. And 8 major chemical markers causing quality differences were screened. CONCLUSION With good reproducibility and stability, this method could provide the reference for the quality evaluation of LHQW．

Objective To explore the approach to establish a temporal lobe epilepsy model via intraperitoneal pilocarpine injection in C57BL/6J mice,and to summarize behavioral indicators predicting successful modeling during the acute phase of epileptic seizures after pilocarpine administration,aiming to offer a practical mice model for future epilepsy research.Methods Thirty C57BL/6J substrain mice(primary subjects)and forty C57BL/6N substrain mice(control subjects)were selected to establish a temporal lobe epilepsy model by inducing seizures through a single intraperitoneal injection of pilocarpine.The mice from the two substrains were each divided into 3 groups,and were injected intraperitoneally with 300 mg/kg,330 mg/kg,or 360 mg/kg of pilocarpine,respectively.Motor seizure behaviors were observed and compared between the two substrains of C57BL/6 mice post pilocarpine injection,and the spontaneous recurrent seizures(SRS)were continuously monitored from the 7th day after injection.On the 28th day post-injection,the mice were euthanized and the histopathological changes in their hippocampi were examined.Results After pilocarpine administration,C57BL/6N mice displayed characteristic motor seizures followed by the onset of status epilepticus(SE).Conversely,C57BL/6J mice showed fewer instances of typical motor seizure behavior and the subsequent SE.Instead,they more often exhibited systemic tremors lasting several seconds to tens of seconds following limb twitching.This behavior is classified as"severe seizure(SS)"in current study.Following intraperitoneal injection of 330 mg/kg and 360 mg/kg pilocarpine,C57BL/6J mice displaying SS during the acute phase of seizure might exhibit SRS after a latency period.The percentage of spontaneous seizures observed in C57BL/6J mice post-modeling(70% )was comparable to that seen in C57BL/6N mice(75% )which developed SRS subsequent to SE.C57BL/6J mice displayed characteristic pathological alterations associated with temporal lobe epilepsy in the hippocampi after 28 d following pilocarpine injection,including increased mossy fiber sprouting and neuronal death.Conclusions When inducing an epilepsy model via intraperitoneal pilocarpine injection in C57BL/6J mice,the behavioral criteria to predict the successful establishment of the model could be either the occurrence of SE or the manifestation of SS.

Cerebrospinal fluid (CSF) laboratory tests are important for diagnosing central nervous system (CNS) diseases. Research on intrathecal immunoglobulin-related indexes has gradually attracted attention. The antibody index, which corrects for the effect of individual blood-brain barrier function on CSF antibody test results, is of great significance in the differential diagnosis, efficacy monitoring and prognostic assessment of CNS diseases. It is expected to become a new index for the diagnosis of CNS diseases. This article reviews the concept of antibody index and the research progress of differential diagnosis and treatment of various CNS diseases in order to provide references for the diagnosis, efficacy monitoring and disease progression assessment of CNS diseases.

Objective:To evaluate the value of Treponema pallidum ( Tp)-specific antibody index in the diagnosis, staging, and typing of neurosyphilis (NS). Methods:Fifty patients diagnosed with NS at the West China Hospital of Sichuan University from March 2020 to December 2022 were recruited as the experimental group, and 50 non-NS syphilis patients were enrolled during the same period as the control group. Cerebrospinal fluid (CSF) and matched serum samples along with clinical data were collected. Clinical and Laboratory Standards Institute (CLSI) EP06-Ed2 file was used to analyze the linear range of electrochemiluminescence immunoassay for Tp-specific antibody detection, and the Tp antibody index was calculate for all subjects. The correlation of Tp antibody index, IgG index, IgG synthesis rate, and albumin quotient with other clinical indicators was evaluated by the coefficient of correlation, and the diagnostic efficacy of these indicators in NS was analyzed using the receiver operating characteristic (ROC) curve. Results:All the deviations from linearity of electrochemiluminescence immunoassay for detecting Tp antibodies in serum and CSF samples were within the allowable deviation from linearity. Compared with the non-NS group, the NS group showed a significant increase in the Tp antibody index ( P<0.05), with no significant differences observed among the groups of different subtypes, severity, or receiving treatment or not ( P>0.05). Tp antibody index, IgG index, and IgG synthesis rate were positively correlated with serum Tp antibody, CSF Tp antibody, trace protein, and nucleated cell count ( P<0.05). Taking 14.99 as the cut-off value for NS diagnosis, the diagnostic sensitivity of Tp antibody index was 74.0% (95%CI: 60.5%-84.1%), and the specificity was 94.0% (95%CI: 83.8%-98.4%), with the area under the ROC curve of 0.897, which was larger than that of albumin quotient, IgG index, and IgG synthesis rate. Conclusions:This study finds a significant increase in the Tp antibody index in NS patients with good diagnostic sensitivity and specificity, providing reference for the diagnosis of NS.

Cerebrospinal fluid (CSF) laboratory tests are important for diagnosing central nervous system (CNS) diseases. Research on intrathecal immunoglobulin-related indexes has gradually attracted attention. The antibody index, which corrects for the effect of individual blood-brain barrier function on CSF antibody test results, is of great significance in the differential diagnosis, efficacy monitoring and prognostic assessment of CNS diseases. It is expected to become a new index for the diagnosis of CNS diseases. This article reviews the concept of antibody index and the research progress of differential diagnosis and treatment of various CNS diseases in order to provide references for the diagnosis, efficacy monitoring and disease progression assessment of CNS diseases.

Objective:To investigate the incidence rate and gene variation of methylmalonic academia (MMA) in Ji′nan city by analyzing biochemical and genetic screening results, and to explore the carrier frequency of MMA-related pathogenic genes in the population in Ji′nan.Methods:The children diagnosed with MMA by tandem mass spectrometry screening in Ji′nan Neonatal Disease Screening Centre from May 2011 to May 2022 were enrolled in this study.Their genetic test results were retrospectively analyzed and summarized.The dried heel blood tablets collected from 6 800 newborns were tested for neonatal gene screening. MMAA, MMAB, MMACHC and MMUT genes in 4 800 cases were detected by high-throughput sequencing+ target area capture technology.Ultra-multiplex polymerase chain reaction+ target gene locus capture technology was used to detect 174 target loci of 8 genes related to MMA in 2 000 cases.The hotspot mutation and related gene carrier rate of MMA were analyzed. Results:A total of 367 452 newborns were screened by tandem mass spectrometry, and 103 cases (56 males and 47 females) were diagnosed with MMA by screening.The estimated incidence of MMA was 1∶3 567.Among the 103 MMA cases, 76 were genetically diagnosed, and 4 gene variants of MMA ( MMAHC, MMUT, MMAA, MMADHC) were identified.A total of 6 800 neonates underwent neonatal genetic screening.Three of them were diagnosed with MMA.About 318 infants carried pathogenic variants of MMA, with a total carrier rate of 4.68%.Specifically, the carrier rates of MMACHC and MMUT gene variants were 3.09%(210/6 800) and 1.43% (97/6 800), respectively. Conclusions:MMA is the most common organic acid metabolism disorder in our country.The incidence and carrier rate of this disease are high in Jinan city.Neonatal genetic screening is an important supplement to neonatal biochemical screening.Carrier screening for MMA-related pathogenic genes is recommended for couples of childbearing age in Jinan.